PROJECT SUMMARY We request funds to purchase a high-resolution and high-sensitivity liquid chromatography- mass spectrometry system for advanced protein and metabolite analysis. This platform is required to significantly enhance capabilities of our Core facility and to meet the needs of NIH- funded investigators for high-resolution, high-mass accuracy and high-sensitivity mass spectrometry, in order to elucidate structures, confirm identities, and perform quantitation of complex molecules such as proteins and metabolites. The instrument will be used in the Central Analytical Laboratory (CAL), which is the MS-based service core facility at the University of Colorado, Boulder. The proposed platform is composed of a high-resolution (up to 240,000 at m/z 200) Thermo Scientific Q Exactive HF mass spectrometer (MS) equipped with a nano ionization source and coupled to a Thermo Scientific Dionex UltiMate 3000 Binary RSLCnano chromatography system for enhanced sensitivity, which is essential for quantitative analysis of low-abundance molecules. The proposed system is needed because our current high-resolution mass spectrometry systems are either non-functional or are in too high demand to fully fulfill the needs of our NIH-funded collaborators at the University of Colorado. Moreover, the proposed instrument is greatly superior to our currently functional high-resolution MS (Thermo LTQ Orbitrap), thus will enable more accurate protein identification and quantitation and greater coverage of protein species. Additionally, the proposed instrument would enable us to expand the capabilities of the Core to include global and targeted metabolic profiling as well as confirmation and identification of novel new metabolites that could potential be used for therapeutic purposes.